| Objectives: By observing of the expression of phenotypic transformation of marker proteins in corpus cavernosum penis of rats to clarify whether the phenotypic modulation of the smooth muscle cell is conducted in the corpus cavernosum penis of DIED rats and the influence of phenotypic transformation;After drug intervention,explore the effect of Schisandrin towards the erectile function of DIED rats and the intrinsic relationship between Schisandrin and smooth muscle cell phenotype transformation proteins.Methods:1 Screen SD rats with DIED: Select 100 healthy male SD rats(200±20g),after 1 week of adaptive feeding,88 rats were randomly injected intraperitoneally with STZ(60mg/kg),the other of them as the normal control.Check the random blood glucose after 72 hours,if the random blood glucose>16.7mmol/L,they were regarded as diabetes mellitus(DM)rats and injected subcutaneously with APO.All rats are fed for 8 weeks,the penile erection of these rats with diabetes is not induced,and accordingly these were regarded as the rats with DIED.Those rats with the blood glucose >16.7mmol/L and with induced penile erection were regarded as the DM rats.2 Grouping for model and the pharmacological intervention: The rats with DIED were randomly divided into four groups as follow,DIEDgroup,low-dose Schisandrin group(5mg/kg/d),middle-dose Schisandrin group(10mg/kg/d)and high-dose Schisandrin group(20mg/kg/d).Besides,NC group and DM group were set.All the ratsof NC group,DM group and DIED group were perfused with 2ml/d normal saline for 8 weeks,and the other disposed groups were conducted the perfusion of medicine intervention for 8 weeks with the corresponding dosage of each.3 After 8 weeks,the SD rats were anesthetic,the nerves of rats’ penises were stimulated electrically,and the ICP,MAP were measured.4 The inferior vena cava blood of rats were collected to measure the level of ALT,AST,ALP and Scr.5 Decapitate the models,clip the penis tissue.Immunohistochemical SP stain the tissue slice as to observe the change of SMA-α from different disposed groups.The protein expression levels of SMA-α and OPN in the other corpus cavernosum penis tissue were determined by applying Western Blot.6 Data analysis: Apply SPSS 23.0 software to conduct the statistical analysis of the acquired data.The measurement data desScription were expressed by((?)±s).Adapt the variance analysis as to conduct the comparison among the normally distributed measured data groups.The multiple comparisons among multi-sample averages were achieved through applying LSD and SNK-q.If the P<0.05,the differences were significant statistically.Result:1 Weight and blood glucose changes of rats from each groups:Compared with NC group,the rat weight before perfusion of the DM group,DIED group,low-dose Schisandrin group,middle-dose Schisandrin group and high-dose Schisandrin group were obviously descending,and the blood glucose was rising obviously(P<0.05).Except for NC group,the comparisons between any two groups were no obvious difference in rat weight and blood glucose(P>0.05).After 8 weeks,compared with NC group,the rat weights of the DM group,DIED group,low-dose Schisandrin group,middle-dose Schisandrin group and high-dose Schisandrin group were obviously descending,and the blood glucose was rising obviously(P<0.05).Except for NC group,the comparisons between any two groups were no obvious difference in rat weight and blood glucose(P>0.05).2 The baseline ICP among all groups had no obvious difference before the electrical stimulation(P>0.05).The MAP among all groups had no obvious difference when the electrical stimulation(P > 0.05).When the electrical stimulation,compared with NC group,the ICP and ICP/MAP of rats from DM and DIED groups were jointly descreasing in sequence(P<0.05).Compared with the DIED group,the ICP and ICP/MAP of rats from DIED and low-dose Schisandrin group had no significant changes(P>0.05),and the ICP and ICP/MAP of rats from middle-dose Schisandrin group and high-dose Schisandrin group were both inscreasing in sequence(P<0.05).3 The concentrations of ALT,AST,ALP and Scr from all groups were relatively the same(P>0.05).4 The immunohistochemical results were shown in the following figure.Compared with NC group,the expressions of SMA-α in DM group and DIED group were descreasing in sequence(P<0.05).Compared with the DIED group,the expressions of SMA-α in low-dose Schisandrin group,middle-dose Schisandrin group and high-dose Schisandrin group were inscreasing in sequence(P<0.05),and were under the expression of NC group.5 Western Blot ResultExpression of protein SMA-α: Compared with NC group,the protein levels of SMA-α in DM and DIED groups were dropping(P<0.05),and the SMA-α protein content of penis tissue of DIED group was less(P<0.05).Compared with the DIED group,the protein contents of SMA-α in each disposed groups were higher than the DIED group’s(P<0.05),the SMA-α protein content of penis tissue of high-dose Schisandrin group was the highest,middle-dose Schisandrin group ranks the second highest place and the lowest SMA-α protein content of penis tissue was from low-dose Schisandrin group.The protein levels of all disposedgroups were lower than NC group’s(P <0.05).Expression of protein OPN: Compared with NC group,the protein levels of OPN in DM and DIED groups were inScreaseing(P<0.05),and the OPN protein content of penis tissue of DIED group was most(P<0.05).Compared with the DIED group,the content of OPN protein in each disposed groups were lower than the DIED group’s(P<0.05),The OPN protein expressions among all disposed groups had no obvious difference(P>0.05).The OPN protein level of penis tissue in DM group was the lowest except for NC group’s(P<0.05).Conclusion:1 It can be predicted that phenotypic modulation of the smooth muscle cell might appear in the penis tissue of rats with DIED,the erectile dysfunction can be caused in line with different phenotypic modulation levels.2 The application of Schisandrin for treatment can improve the erection function of rats with DIED.The impact of Schisandrin for improving the erection function is in a dose-dependent manner.The more effective the optimizing impact of high-dose group is the strongest.3 The Schisandrin has not exert negative influence to the hepatorenal and kidney function of rats with DIED.4 Schisandrin may affect the expression of phenotypic transforming protein in smooth muscle cells of corpus cavernosum penis,but it is no dose-dependent. |
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